High temperature treatment induced production of unreduced 2n pollen in Camellia oleifera

Unreduced gametes through chromosome doubling play a major role in the process of plant polyploidization.Our previous work confirmed that Camellia oleifera can produce natural 2n pollen,and it is possible to induce the 2n pollen formation by high temperature treatment.This study focused on the optimization of the 2n pollen induction technique and the mechanisms of high temperature-induced2n pollen formation in C.oleifera.We found that the optimal protocol for inducing 2n pollen via high temperature was to perform 45℃with4 h at the prophaseⅠstage of the pollen mother cells(PMCs).Meanwhile,high temperature significantly decreased the yield and fertility of2n pollen.Through the observation of meiosis,abnormal chromosome and cytological behaviour was discovered under high-temperature treatment,and we confirmed that the formation of 2n pollen is caused by abnormal cell plate.Based on weighted gene co-expression network analysis,fifteen hub genes related to cell cycle control were identified.After male flower buds were exposed to heat shock,polygalacturonase gene(CoPGX3)was significantly upregulated.We inferred that high temperature causes the CoPGX3 gene to be overexpressed and that CoPGX3 is redistributed into the cytosol where it degrades cytoplasmic pectin,which leads to an abnormal cell plate.Furthermore,abnormal cytokinesis resulted in the formation of dyads and triads,and PMCs divided to produce 2n pollen.Our findings provide new insights into the mechanism of 2n pollen induced by high temperature in a woody plant and lay a foundation for further ploidy breeding of C.oleifera.

Effect of different drying methods on the amino acids,α-dicarbonyls and volatile compounds of rape bee pollen

The significant demand for high quality food has motivated us to adopt appropriate processing methods to improve the food nutritional quality and flavors.In this study,the effects of five drying methods,namely,pulsed vacuum drying(PVD),freeze drying(FD),infrared drying(IRD),hot-air drying(HAD)and sun drying(SD)on free amino acids(FAAs),α-dicarbonyl compounds(α-DCs)and volatile compounds(VOCs)in rape bee pollen(RBP)were determined.The results showed that FD significantly released the essential amino acids(EAAs)compared with fresh samples while SD caused the highest loss.Glucosone was the dominantα-DCs in RBP and the highest loss was observed after PVD.Aldehydes were the dominant volatiles of RBP and SD samples contained more new volatile substances(especially aldehydes)than the other four drying methods.Comprehensively,FD and PVD would be potential methods to effectively reduce the quality deterioration of RBP in the drying process.

GAMYB transcription factor LoMYB65 from lily plays a vital role in pollen development

Lily(Lilium spp.)is an important horticultural crop,but its use is limited due to serious pollen contamination problems.There are many studies on pollen development in model plants,but few on flower crops such as lilies.Gibberellin(GA)is a large class of hormones and plays an important role in plant vegetative growth and reproductive development.GAMYB is a group of the R2R3-MYB family upregulated by gibberellin,and plays an important role in anther development.Here,we isolated a novel GAMYB,named LoMYB65,from lily,which was closely related to the AtMYB65 and AtMYB33 in Arabidopsis.Fluorescence quantitative PCR results showed that LoMYB65 was mainly expressed in lily anthers.LoMYB65 could be activated by 288μmol·L^(-1)GA3treatment and the LoMYB65 protein was located in the nucleus and cytoplasm,and had transactivation in yeast and tobacco leaf cells.The conserved motif within 226 amino acids of the C-terminal of LoMYB65 contributed to its transactivation.Overexpression of LoMYB65 caused dwarf phenotype,unnormal tapetum development,less seeds of siliques in transgenic Arabidopsis plants,the transgenic plants showed partly male sterile.Simultaneously,silencing of LoMYB65 with VIGS(Virus Induced Gene Silencing)in lily anthers caused unnormal pollen development and reduced the pollen amount.Overexpression of LoMYB65 in Arabidopsis and silencing of LoMYB65 in lily resulted in decreased pollen counts,so we speculate that LoMYB65 may be dose-dependent.Overall,these findings suggest that LoMYB65 may play an important role in anther development and pollen formation in lily.LoMYB65 may provide a useful candidate gene for pollenless breeding of lily.

Effect of wall-disruption on nutrient composition and in vitro digestion of camellia and lotus bee pollens

The nutrient digestion,absorption and biological activity of bee pollen may be limited due to the complex pollen wall.Here,the effect of superfine grinding technology on the release of nutrients from bee pollen were investigated,and their antioxidant activities and in vitro digestion were explored in this study.Results showed that the content of nutrients in bee pollen increased after wall disruption.Among them,fat content increased by 22.55%-8.31%,protein content increased by 0.54%-4.91%,starch content increased by 36.31%-48.64%,soluble sugar content increased by 20.57%-29.67%,total phenolic acid content increased by 11.73%-86.98%and total flavonoids content increased by 14.29%-24.79%.At the same time,the antioxidant activity increased by 14.84%-46.00%.Furthermore,the active components such as phenolic compounds in the wall-disruption bee pollen were more readily to be released during the in vitro digestion,and easier to be absorbed because of their higher bioaccessibility.Antioxidant activities during in vitro digestion were also improved in walldisruption bee pollen.These findings provide evidence that bee pollen wall disruption was suggested,thus,it is more conducive to exerting the value of bee pollen in functional foods.

Allergen degradation of bee pollen by lactic acid bacteria fermentation and its alleviatory effects on allergic reactions in BALB/c mice

Food allergy as a global health problem threatens food industry.Bee pollen(BP)is a typical food with allergenic potentials,although it performs various nutritional/pharmacological functions to humans.In this study,lactic acid bacteria(LAB)were used to ferment Brassica napus BP for alleviating its allergenicity.Four novel allergens(glutaredoxin,oleosin-B2,catalase and lipase)were identified with significant decreases in LAB-fermented BP(FBP)than natural BP by proteomics.Meanwhile,metabolomics analysis showed significant increases of 28 characteristic oligopeptides and amino acids in FBP versus BP,indicating the degradation of LAB on allergens.Moreover,FBP showed alleviatory effects in BALB/c mice,which relieved pathological symptoms and lowered production of allergic mediators.Microbial high-throughput sequencing analysis showed that FBP could regulate gut microbiota and metabolism to strengthen immunity,which were closely correlated with the alleviation of allergic reactivity.These findings could contribute to the development and utilization of hypoallergenic BP products.

DIA-based proteome profiling with PRM verification reveals the involvement of ER-associated protein processing in pollen abortion in Ogura CMS cabbage

Ogura cytoplasmic male sterility(Ogura CMS)is extensively applied in hybrid seed production in cruciferous crops.However,the posttranscriptional molecular basis of Ogura CMS in cruciferous crops remains elusive.Here,a data-independent acquisition-based proteomic approach coupled with a parallel reaction monitoring-based targeted proteomic assay was used to analyze the proteome dynamics of Ogura CMS cabbage line RM and its maintainer line RF during floral bud development to obtain insights into the mechanism underlying Ogura CMS in cruciferous crops.A total of 9162 proteins corresponding to 61464 peptides were identified in RM and RF floral buds.The proteomic fluctuation of RM was weaker than that of RF.Differences in protein expression between RM and RF gradually enlarged with floral bud development.Fifteen continually up-regulated and eight continually down-regulated proteins were found in RM relative to RF throughout floral bud development.Differentially expressed proteins between RM and RF during floral bud development were implicated in the endoplasmic reticulum(ER)-associated protein processing pathway,in which most of them exhibited down-regulated expression in RM.These data suggest that ER-associated protein processing may be involved in pollen abortion in Ogura CMS cabbage by inhibiting the expression of critical factors.Our findings not only deepen the understanding of the molecular mechanisms of Ogura CMS in cruciferous crops but also provide better guidance for applying Ogura CMS in the hybrid breeding of cruciferous crops.

Identification and functional analysis of arabinogalactan protein expressed in pear pollen tubes

Arabinogalactan proteins(AGPs)are widely distributed in the plant kingdom and play a vital role during the process of plant sexual reproduction.In this study,we performed a comprehensive identification of the PbrAGPs expressed in pear pollen and further explored their influences on pollen tube growth.Among the 187 PbrAGPs that were found to be expressed in pear pollen tubes,38 PbrAGPs were specifically expressed in pollen according to the RNA-seq data.The PbrAGPs were divided into two groups of highly expressed and specifically expressed in pear pollen.We further tested their expression patterns using RT-PCR and RT-qPCR.Most of the PbrAGPs were expressed in multiple tissues and their expression levels were consistent with reads per kilobase per million map reads(RPKM)values during pollen tube growth,implying that PbrAGPs might be involved in the regulation of pear pollen tube growth.We also constructed phylogenetic trees to identify the functional genes in pear pollen tube growth.Therefore,19 PbrAGPs(PbrAGP1 to PbrAGP19)were selected to test their influences on pollen tube growth.Recombinant proteins of the 19 PbrAGP-His were purified and used to treat pear pollen,and 11 of the PbrAGP-His recombinant proteins could promote pear pollen tube growth.Additionally,pollen tube growth was inhibited when the expression levels of PbrAGP1 and PbrAGP5 were knocked down using an antisense oligonucleotide assay.PbrAGP1 and PbrAGP5 were localized in the plasma membrane and might not alter the distribution of pectin in the pollen tube.In summary,this study identified the PbrAGPs expressed in pear pollen and lays the foundation for further exploring their functions in pollen tube growth.

Modern pollen assemblages and their relationships with vegetation and climate on the northern slopes of the Tianshan Mountains, Xinjiang, China

The reconstruction of paleovegetation and paleoclimate requires an understanding of the relationships between surface pollen assemblages and modern vegetation and climate.Here,we analyzed the characteristics of surface pollen assemblages across different vegetation zones in the Tianshan Mountains.Using surface pollen analysis and vegetation sample surveys at 75 sites on the northern slopes of the Tianshan Mountains,we determined the correlation between the percentage of dominant pollen types and the corresponding vegetation cover.Redundancy analysis was used to investigate the relationships between surface pollen assemblages and environmental factors.Our results show that the Tianshan Mountains contain several distinct ecological regions,which can be divided into five main vegetation zones from low to high altitudes:mountain desert zone(Hutubi County(HTB):500-1300 m;Qitai County(QT):1000-1600 m),mountain steppe zone(HTB:1400-1600 m;QT:1650-1800 m),mountain forest zone(HTB:1650-2525 m;QT:1850-2450 m),subalpine meadow zone(HTB:2550-2600 m;QT:2500-2600 m),and alpine mat vegetation zone(HTB:2625-2700 m;QT:2625-2750 m).The surface pollen assemblages of different vegetation zones can accurately reflect the characteristics of the mountainous vegetation patterns on the northern slopes of the Tianshan Mountains when excluding the widespread occurrence of Chenopodiaceae,Artemisia,and Picea pollen.Both average annual precipitation(P_(ann))and annual average temperature(T_(ann))affect the distribution of surface pollen assemblages.Moreover,P_(ann) is the primary environmental factor affecting surface pollen assemblages in this region.A significant correlation exists between the pollen percentage and vegetation cover of Picea,Chenopodiaceae,Artemisia,and Asteraceae.Moreover,Picea,Chenopodiaceae,and Artemisia pollen are over-represented compared with their corresponding vegetation cover.The Asteraceae pollen percentage roughly reflects the distribution of a species within the local vegetation.These results have important implications for enhancing our understanding of the relationship between surface pollen assemblages and modern vegetation and climate.

First Report of a Successful Development of Yam Hybrids(Dioscorea alata L.)from Lyophilized and Long-Term Stored Pollens

Various biological constraints including erratic and asynchronous flowering between male and female plants hinder successful hybrid development and genetic gains in greater yam breeding programs.Therefore,pollen storage has gained much attention to facilitate artificial pollinations and increase the genetic gains.This 4-year study aimed at developing a practical long-term pollen storage technique for the successful development of yam hybrids.Fresh pollens were collected from two Dioscorea alata males,then lyophilized(two lyophilization treatments were applied),followed by storage at room temperature(24℃–25℃)for 12 months.Moreover,the lyophilized and stored pollens were tested for viability by crossing with four female varieties.Our results showed that lyophilization is effective for achieving viable pollens after 12 months of storage.Treatment 1(48 h drying)showed higher pollen germination and fertility rates than Treatment 2(72 h drying).Although we observed a reduction in viability of lyophilized pollens after 12 months of storage,we generated hybrid seedlings with success rates from 12%to 21%compared to 21%–31%when using fresh pollens.Paternity testing based on molecular genotyping confirmed the hybrid status of the obtained seedlings,which grew well in a greenhouse.Lyophilization is a practical approach for a long-term storage of greater yam pollen samples.This protocol will positively impact yam breeding programs particularly in developing countries.

Relationships between modern pollen and vegetation and climate on the eastern Tibetan Plateau

The Tibetan Plateau ecosystem is fragile and sensitive to climate change. Understanding the relationships between modern pollen and the vegetation and climate of the region is critical for the evaluation of ecological processes.Here, we explore modern pollen assemblages of typical land-cover types at a large spatial scale by analyzing 36surface samples from the southeastern Tibetan Plateau, supplemented by typical desert, desert-steppe, and steppe meadow transition data selected from the Chinese Surface Pollen Database, giving a total of 75 samples. We used redundancy analysis(RDA) to explore the responses of vegetation in the assemblages to regional climate. Our results show that pollen assemblages generally reflect the vegetation composition: assemblages from alpine meadow samples are dominated by Cyperaceae, Asteraceae, Rosaceae, and Polygonaceae;alpine shrublands mainly comprise Fabaceae, Rosaceae, Ericaceae, and Quercus(Q. spinosa);and coniferous forest surface samples mainly comprise Picea, Abies, Pinus, and Betulaceae. Our RDA shows that mean annual precipitation(MAP) is the main meteorological factor affecting the pollen assemblage and vegetation type;MAP positively correlates with percentages of Cyperaceae, Poaceae, Rosaceae, and Asteraceae, and negatively correlates with percentages of Chenopodiaceae, Ephedraceae, Nitraria, and Tamaricaceae. The ratio of Artemisia to Chenopodiaceae is a useful indicator to distinguish temperate desert from other land-cover types on the Tibetan Plateau, while the ratio of Cyperaceae + Asteraceae to Artemisia + Chenopodiaceae can be used to distinguish arid desert from other landcover types, and may provide a useful altitude index for the eastern Tibetan Plateau.

The morphological diversity of pollen in the genus Gossypium

Background Plant pollen has diverse morphological characteristics that can be consistently passed down from generation to generation.Information on pollen morphology is thus immensely important for plant classification and identification.In the genus Gossypium,however,in-depth research on pollen morphology is lacking,with only few reports on limited cotton species.To evaluate the diversity of pollen in Gossypium,we therefore conducted a comprehensive analysis of the pollen morphology of 33 cotton species and varieties using scanning electron microscopy.Results The 33 analyzed cotton samples exhibited common pollen morphological features,including spherical shapes,radial symmetry,echination,panporation,and operculation,while the pollen size,spine shape,spine density and length showed distinctive features.Pollen size varied significantly among species,with diameters ranging from62.43 μm in G.harknessii to 103.41 μm in G.barbadense.The exine had an echinate sculptural texture,and spines were mostly conical or sharply conical but occasionally rod-like.Spine density varied from 173 in G.incanum to 54 in G.gossypioides,while spine length ranged from 3.53 μm in G.herbaceum to 9.47 μm in G.barbadense.In addition,the 33cotton species and varieties were grouped at a genetic distance of 3.83 into three clusters.Cluster Ⅰ comprised five allotetraploid AD-genome cotton species,four D-genome species,and one K-genome species.Cluster Ⅱ included 13diploid species from A,B,D,E,and G genomes,whereas Cluster Ⅲ only consisted one E-genome species G.incanum.Conclusions Although pollen characteristics alone are not enough to resolve taxonomic and systematic relationships within the genus Gossypium,our results add to knowledge on palynomorphology and contribute to phenological information on these taxa.Our findings should aid future systematic and phylogenetic studies of the Gossypium genus.

Current Status of Research on Pollen Vitality and Stigma Receptivity of Wuren Walnut(Juglans sigillata)in Guizhou Province

Wuren walnut is a unique germplasm resource of Juglans sigillata in Guizhou Province,named after its purple and black seed coat.It is a typical medicinal and edible plant with high nutritional value and good taste compared with ordinary walnut.The color of its inner seed coat is different from that of ordinary walnut,which is mostly light yellow.Wuren walnut has great market potential for development.To lay a theoretical foundation for the future breeding of improved varieties,pollination tree configuration,and hybrid breeding of Wuren walnut in Guizhou Province,this paper summarized the current research status of pollen vitality and stigma acceptability of Guizhou Wuren walnut in recent years,and looked forward to future research and application.

Preliminary Study on in Vitro Germination of Yangmei Pollen

[Objectives]To explore the conditions for in vitro culture of Yangmei pollen.[Methods]Experiments were conducted on the germination characteristics of three types of Yangmei pollen using in vitro culture and germination method.[Results]The suitable medium ratio for the germination of Yangmei pollen was 10%sucrose+0.01%borax+1%agar;the cultivation temperature of 30℃was more suitable for the germination of Yangmei pollen than 25℃;through analysis of variance,among the three types of Yangmei pollen,pollen of male 1 had the strongest viability,and it was the better pollination type.[Conclusions]The research could provide certain basis for introduction and cultivation of Yangmei and improving hybrid breeding effect.

Plant Pollen Grains:A Move Towards Green Drug and Vaccine Delivery Systems

Pollen grains and plant spores have emerged as innovative biomaterials for various applications such as drug/vaccine delivery,catalyst support,and the removal of heavy metals.The natural microcapsules comprising spore shells and pollen grain are designed for protecting the genetic materials of plants from exterior impairments.Two layers make up the shell,the outer layer(exine)that comprised largely of sporopollenin,and the inner layer(intine)that built chiefly of cellulose.These microcapsule shells,namely hollow sporopollenin exine capsules have some salient features such as homogeneity in size,non-toxic nature,resilience to both alkalis and acids,and the potential to withstand at elevated temperatures;they have displayed promising potential for the microencapsulation and the controlled drug delivery/release.The important attribute of mucoadhesion to intestinal tissues can prolong the interaction of sporopollenin with the intestinal mucosa directing to an augmented effectiveness of nutraceutical or drug delivery.Here,current trends and prospects related to the application of plant pollen grains for the delivery of vaccines and drugs and vaccine are discussed.

Less and shrunken pollen 1(LSP1) encodes a member of the ABC transporter family required for pollen wall development in rice(Oryza sativa L.)

Pollen fertility is an agronomic trait that strongly influences rice yield. Recent studies have revealed that the development of the pollen wall is required for pollen fertility and is regulated by several genes. However, the mechanisms underlying pollen and pollen wall development in rice remain largely unknown. In the present study, a point mutation in a gene on chromosome 1 was identified that resulted in the production of less and shrunken pollen(LSP) and led to defects in pollen wall formation. This gene was named LSP1 and was found to encode a member of the adenosine triphosphate-binding cassette(ABC)transporter G subfamily, OsABCG3. Two other loss-of-function mutants of LSP1/OsABCG3,generated using CRISPR/Cas9 technology, showed the same male sterile phenotype. The LSP1/OsABCG3 gene showed a spatio-temporal expression pattern in the developing anthers, and is an ortholog of the Arabidopsis genes At ABCG1 and At ABCG16, which play an important role in pollen wall development. Mutation of LSP1/OsABCG3 affected the expression of several genes involved in pollen and pollen wall formation. These results suggest that LSP1/OsABCG3 is critical for normal pollen fertility and shed light on the molecular mechanisms underlying rice pollen wall development.

Study on the Pollen Thermo-Sensitive and Stigma Exsertion Male Sterile Line Tomato (Lycopersicon esculentum Mill.) cv. Da107

To accelerate the breeding process, male sterile line is used to leave out the troublesome procedures of the artificial emasculation in tomato breeding. However, the fertility of the pollen thermo-sensitivity male sterile line (PTMSL) and the stigma exsertion male sterile line (SEMSL) are affected easily by the environments when used alone. The trial materials were Da107 and the control was First. This study was conducted to create a new male sterile line of tomato characterized by pollen thermo-sensitivity and long style by genic recombination through the hybridizing of the PTMSL and SEMSL. Research on the statistics of the pollen germination rate, the contamination rate in F1 and the flower organics indicated that Da107 was an ideal TS and SE male sterile line with the sterility of 95%, as well as it also could be used as fertile line at low temperature. Meanwhile, the results showed that hybrid-seed contamination risk with selfed seeds from residual fertility in Da107 was low.

Influence of Storage Time on Pollen Traits in Actinidia eriantha

We examined the influence of storage time on germinability and tube growth of freeze stored pollens collected from 25 wild male plants in Actinidia eriantha variety. Pollens were stored in freezer at - 20°C for six months and one year periods to determine changing at germinability in time. In vitro germination was conducted in certain cultural medium defined for Actinidia genus. The results showed that the germination percentages and tube lengths of genotypes decreased at the end of storage period. MH22, MH45, MH47, MH56, MH67, MH70, MH71, MH72, MH74, MH55 and MH61 genotypes were evaluated as vigor genotypes, because they maintained their viability and germination capability displaying statistically insignificant decreasing although their tube lengths significantly decreased except MH67. This investigation provided to determine some robust wild male germplasm resources in A. eriantha in point of durability of pollens against long term conservation for using at future pollination and breeding programs.

Low soil temperature and drought stress conditions at flowering stage affect physiology and pollen traits of rice

Low temperature and drought stress are the major constraints in rice productivity worldwide. This study investigated the influence of low soil temperature and/or drought stress on physiology and pollen traits of two rice genotypes viz., Guinongzhan and Yueza 763 at flowering stage. The experiment included four treatments, i.e., under the greenhouse natural growth conditions (UC) taken as control, drought stress (DS), the soil water potential was kept at ?0.035 to ?0.045 MPa (DS), low soil temperature (LT) maintained at 19 to 21℃, combined LT and DS (LT+DS, LD). Results showed that LT, DS, and LD substantially reduced net photosynthetic rate (Pn) and the maximal photochemical efficiency of PSII (Fv/Fm), whilst transpiration rate (Tr) was markedly enhanced by under LT in both rice genotypes. The malondialdehyde (MDA) contents were enhanced under LT, DS, and LD in Guinongzhan, whilst the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were regulated by LT, DS, and LD in both rice genotypes. Furthermore, anther dehiscence rate, pollen numbers on stigma, pollen viability and pollen germination rate, and anther starch contents were obviously reduced under LT and DS of both rice genotypes. Stress conditions substantially reduced the yield and yield components, i.e., effective panicles, seed set percentage, grain/panicles, 1000-grain weight, and grain yield of both genotypes and the effects were more apparent in Guinongzhan than those in Yueza 763 whilst combined LT and DS proved more damaging than individual stress.

Quantitative Trait Loci Associated with Pollen Fertility under High Temperature Stress at Flowering Stage in Rice (Oryza sativa)

High temperature stress (HTS), an increasingly important problem in rice production, significantly reduces rice yield by reducing pollen fertility and seed setting rate. Breeding rice varieties with tolerance to HTS at the flowering stage is therefore essential for maintaining rice production as the climate continues to become warm. In this study, two quantitative trait loci (QTLs) underlying tolerance to HTS were identified using recombinant inbred lines derived from a cross between an HTS-tolerant rice cultivar 996 and a sensitive cultivar 4628. Pollen fertility was used as a heat-tolerance indicator for the lines subjected to HTS at the flowering stage in field experiments. Two QTLs that affected pollen fertility, qPF4 and qPF6, were detected between RM5687 and RM471 on chromosome 4, and between RM190 and RM225 on chromosome 6, by using the composite interval mapping (CIM) analysis. The two QTLs explained 15.1% and 9.31% of the total phenotypic variation in pollen fertility, and increased the pollen fertility of the plants subjected to HTS by 7.15% and 5.25%, respectively. The positive additive effects of the two QTLs were derived from the 996 alleles. The two major QTLs identified would be useful for further fine mapping and cloning of these genes and for molecular marker-assistant breeding of heat-tolerant rice varieties.

Rice False Smut Fungus, <i>Ustilaginoidea virens</i>, Inhibits Pollen Germination and Degrades the Integuments of Rice Ovule

Ustilaginoidea virens is a flower-infecting fungus that forms false smut balls in rice panicle. Rice false smut has long been considered a minor disease, but recently it occurred frequently and emerged as a major disease in rice production. In vitro co-cultivation of U. virens strain with young rice panicles showed that U. virens enters inside of spikelets from the apex and then grows downward to infect floral organs. In response to U. virens infection, rice host exhibits elevated ROS accumulation and enhanced callose deposition. The secreted compounds of U. virens can suppress rice pollen germination. Examination of sectioning slides of freshly collected smut balls demonstrated that both pistil and stamens of rice flower are infected by U. virens, hyphae degraded the contents of the pollen cells, and also invaded the filaments. In addition, U. virens entered rice ovary through the thin-walled papillary cells of the stigma, then decomposed the integuments and infected the ovary. The invaded pathogen could not penetrate the epidermis and other layers of the ovary. Transverse section of the pedicel just below the smut balls showed that there were no fungal hyphae observed in the vascular bundles of the pedicel, implicating that U. virens is not a systemic flower-infecting fungus.